This document summarizes the micropropagation of sugarcane through tissue culture. It discusses the importance of sugarcane as a crop and the need for tissue culture to rapidly produce disease-free, new varieties. The key steps of sugarcane tissue culture protocols are explained in brief, including explant source, medium, callus induction, shoot regeneration, rooting, environmental conditions, and acclimatization. The optimal conditions and plant growth regulator concentrations for each step are outlined. The overall goal of the protocols described is to efficiently regenerate new sugarcane plants through biotechnology that can then be commercialized.

1. Micropropogation of saccharum
(sugarcane)
By: Maira sultan

2. Introduction
• Grass family (Poaceae)
• Main product = sucrose fig. 1: sugarcane feilds
• World's largest crop by production quantity
• Brazil =largest producer
• Pakistan =Punjab = 60 - 65 %

3. Tissue culture needs
• Rapid, large-scale production
• Disease-free
• Newly developed varieties
• Speed up the breeding
• Commercialization process

4. Tissue culture Protocols
1. Explant source
2. Medium and condition
3. Callus induction
4. Shoot regeneration
5. Rooting medium
6. Environmental condition fig.2: invitro cultures
7. Acclimatization

5. Explant source
• Healthy young meristems
• Removing leaf sheath
• Cut into 1.0 -1.5 cm length
• Washed for 20-30 minutes
• Alcohol (70 %) for 30 seconds
fig.3: young sugarcane plants

6. medium and conditions
• Semisolid basal MS medium
• Concentrations and combinations of different
PGRs

7. Callus induction
• range of concentration of auxins
• 2,4-D (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
• IBA (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
• NAA (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
Fig.4: callus formation in sugarcane

8. Shoot regeneration
• White friable calli
• Cultured on MS medium
• Combinations and concentrations of:
BAP,
Kinetin (0.5- 4.0mg/l)
IBA, NAA (0.1-1.0mg/l)
• Multiple shoot regeneration individually or in
combination

9. Rooting
• Half-strength (1/2 MS)
• IBA
• NAA
• IAA (0.5-3.0mg/l)
fig.5: roots of sugarcane

10. Environmental conditions
• pH = 5.8 before gelling
• Photoperiod = 16h
• Temperature = 25 ± 3 C
• Relative humidity = 70-80%

11. Acclimatization
• Culture medium
• plastic trays
• 1/8 MS basal salt
solution
• After 30 days transplanted in to
field condition

12. Results
• Callus Induction:
• Regeneration of Micro shoots:

13. Thank you